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NCJRS Abstract

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NCJ Number: 229267 Find in a Library
Title: Fast Multiplexed Polymerase Chain Reaction for Conventional and Microfluidic Short Tandem Repeat Analysis
Journal: Journal of Forensic Sciences  Volume:54  Issue:6  Dated:November 2009  Pages:1287-1296
Author(s): Heidi Giese, Ph.D.; Roger Lam, M.Sc.; Richard Selden, M.D., Ph.D.; Eugene Tan, Ph.D.
Date Published: November 2009
Page Count: 10
Type: Report (Study/Research)
Format: Article
Language: English
Country: United States of America
Annotation: This paper reports for the first time the successful amplification of full DNA profiles that consist of either 9 or 15 short tandem repeat (STR) loci and amelogenine in both tube reactions and 16-sample microfluidic biochip reactions in approximately 17 minutes.
Abstract: The protocols and instrumentation presented enable significant reductions in the time required to generate full profiles that satisfy interpretation guidelines for STR analysis. The time required for STR amplification is determined by the temperature ramp rates of the thermal cycler, the components of the reaction mix, and the properties of the reaction vessel. Multiplex amplifications in microfluidic biochip-based and conventional tube-based thermal cyclers have been demonstrated in 17.3 and 19 minutes, respectively. Optimized 28-cycle amplification protocols generated alleles with signal strengths above calling thresholds, heterozygous peak height ratios of greater than 0.65, and incomplete nontemplate nucleotide addition and stutter of less than 15 percent. Full CODIS-compatible profiles were generated using the Profiler PlusID, COfiler and Identifiler primer sets, PCR peresis system, Genebench-FXTM. This fast multiplex PCR approach has the potential to reduce process time and cost for STR analysis and enables development of a fully integrated microfluidic forensic DNA analysis system. The descriptions of methods and materials address the custom thermal cycler and microfluidic biochip, instrumentation and temperature profiles, reaction mix component and cycling conditions for PCR, multiplex PCR with other STR typing kits, reproducibility, sensitivity, STR separation and detection instrumentation, and data analysis. 2 tables, 7 figures, and 35 references
Main Term(s): Criminology
Index Term(s): Cost effectiveness analysis; DNA fingerprinting; Efficiency; Forensic sciences; Investigative techniques
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